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Reproduction is an important life process in insects; however, few studies have attempted to demonstrate the association between reproductive activity and energy metabolism. recent decades, reproductive roles have garnered great attention, and many interesting results have been generated1,2,3,4. In insects, such as tephritid fruit flies, nutrient metabolism has a critical effect on male sexual performance5. For example, nutrient reserves can regulate male participation in leks, as observed in the Mediterranean fruit fly (Wiedemann)6. In the Mediterranean fruit fly, manuka oil can significantly increase the mating success of both wild males and mass-reared, sterile males, and -copaene is the key regulator7. Furthermore, the male diet can affect male mating success Inulin manufacture and longevity as well as female remating8. Reproductive systems can have important ecological and evolutionary implications. Changes in reproductive roles can affect population density9 and the intensity of sexual selection10. In honey bees ((Hendel) (Diptera: Tephritidae) is a notorious pest worldwide that damages many commercial fruits16,17 and a wide range of other agricultural products18. By destroying the marketability of fruit products, this pest has caused dramatic financial loss to orchard crops18. has received considerable attention, and its mating behavior has been investigated for decades19,20. However, these studies have focused on chemical (e.g., pheromones) factors and mating behavior and not on the association between changes in the reproductive activity and energy metabolism. To identify the male mating molecular mechanism, we developed a large-scale oligonucleotide microarray for and investigated the transcriptome profiles in males. males need to take up nutrition for approximately one week Inulin manufacture before mating, and at the end of each mating activity, the male must undergo a complementary nutrition stage prior to another mating activity5,6,15 (Figure 1). We therefore investigated how mating competitiveness differences gradually progress over time and identified pathways, genes and substrates that can be used to determine the molecular and genetic bases involved in mating competitiveness changes in males. Furthermore, we identified reproductive changes through mating competition tests and metabolic changes during different reproductive statuses. Figure 1 Fruit fly mating process timeline5,6,15. Results Evaluation of transcriptome data The error rate for the base positions in the reference transcriptome was less than 1%, which was considered an acceptable threshold (observe Supplementary Physique S1 on-line). Moreover, the base content material distribution demonstrates the G and C or perhaps a and T levels were equivalent, and horizontal lines were observed during the sequencing process, except for a number of bases at the beginning positions (observe Supplementary Physique S1 on-line). After quality control, 26,589,907 clean reads out of 27,364,337 natural reads were acquired for pair-end sequencing. We generated 58,009 transcripts and 33,314 unigenes for which the N50 was 1,849 (observe Supplementary Physique S2 online, Table 1). Table ZPK 1 Size distribution for the put together sequences Functional annotation of the transcriptome Through BLAST searches in the seven indicated databases, 33,314 unigenes were successfully annotated; most Inulin manufacture of the unigenes (15,395) that were annotated were from the non-redundant (NR) database, whereas the fewest (3,864) were from KEGG orthology (KO) (Table 2). In the Gene Ontology (Proceed) database, the top three Proceed terms were cellular process, binding process and metabolic process, for which 8,302, 7,620 and 7,171 genes were annotated, respectively (observe Supplementary Physique S3 online). In clusters of orthologous groups of proteins (KOG), 7,640 annotated genes were assigned to 26 organizations; the (R) general practical prediction only; (T) signal transduction; and (O) post-translational modification, protein turnover and chaperone organizations contained probably the most annotated genes (1,530, 1,163 and 694 genes, respectively) (observe Supplementary Physique S4 on-line). In KEGG, 3,864 genes were annotated with pathways; most of the genes were annotated with the signal transduction; translation; and folding, sorting and degradation groups (622, 497 and 356 genes, respectively) (observe Supplementary Physique S5 on-line). The transcriptome data of males was submitted to TSA database in NCBI (distribution ID: SUB741296). Table 2 Unigene annotation in the databases Inulin manufacture Quality assessment of the sequencing data from males with different reproductive statuses The base error rate in each sample was less than 0.08%, and the rates for the first six positions.